Tibialis anterior muscle samples were washed in phosphate-buffered saline (PBS, Bio-Optica, Milano, Italy), fixed in 10% buffered-formalin (Bio-Optica, Milan, Italy) for 24 h at room temperature. Afterwards, the samples were dehydrated in graded ethanol (Bio-Optica, Milan, Italy), cleaned in xylene (Bio-Optica, Milan, Italy) and paraffin-embedded (Bio-Optica, Milan, Italy), being careful to preserve the desired anatomical orientation. Slides of 5 μm thickness were cut from the obtained paraffin blocks and hematoxylin and eosin-stained (H&E, Bio-Optica, Milan, Italy) following a protocol described elsewhere [59 (link)]. The samples were then examined in triplicate for morphological evaluation with a Zeiss Axioplan light microscope (Carl Zeiss, Oberkochen, Germany) and by a digital camera (AxioCam MRc5, Carl Zeiss, Oberkochen, Germany), used to take images.
Graded ethanol
Manufactured by Bio-Optica
Sourced in Italy
Graded ethanol is a laboratory reagent used for various applications in histology and cell biology. It is a series of ethanol solutions with different concentrations, typically ranging from 70% to 100%. These graded ethanol solutions are used for the dehydration and clearing of tissue samples during the specimen preparation process.
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2 protocols using graded ethanol
1
Histomorphometric Analysis of Tibialis Anterior Muscle
2
Histological Evaluation of Cartilage Repair
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Cartilage samples were washed in phosphate-buffered saline (PBS, Bio-Optica, Milano, Italy), fixed in 10% buffered-formalin (Bio-Optica, Milan, Italy) for 24 h at room temperature. Afterwards, the samples were dehydrated in graded ethanol (Bio-Optica, Milan, Italy), cleaned in xylene (Bio-Optica, Milan, Italy) and paraffin-embedded (Bio-Optica, Milan, Italy), being careful to preserve the desired anatomical orientation. For the general evaluation of the morphological structure of the cartilage, the slides of 4–5 µm thickness were cut from the obtained paraffin blocks and haematoxylin and eosin-stained (H&E; Bio-Optica, Milan, Italy) as previously described [33 (link)]. The samples were then examined with a Zeiss Axioplan light microscope (Carl Zeiss, Oberkochen, Germany) and by a digital camera (AxioCam MRc5, Carl Zeiss), used to take images.
For qualitative histological analysis the following parameters were analysed:
For qualitative histological analysis the following parameters were analysed:
The type of repaired tissue on the lesion surface (cartilaginous, fibrous or calcified);
Capability of the collagen I-based scaffold to recruit host cells and promote cartilaginous matrix deposition;
The scaffold biocompatibility and reabsorption of the collagen I-based scaffold.
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